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Related Experiment Videos

Time-resolved fluorescence anisotropy imaging applied to live cells.

Klaus Suhling1, Jan Siegel, Peter M P Lanigan

  • 1Departments of Physics, Biological Sciences and Chemistry, Imperial College London, London SW7 2BW, UK. k.suhling@imperial.ac.uk

Optics Letters
|March 24, 2004
PubMed
Summary

We developed a new imaging system to measure fluorescence lifetime and rotational correlation time. This technique quantifies solvent viscosity and probes microviscosity in live cells.

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Area of Science:

  • Biophysics
  • Optical Imaging
  • Fluorescence Spectroscopy

Background:

  • Fluorescence lifetime and rotational correlation time provide insights into molecular environments.
  • Quantifying microviscosity in biological systems is crucial for understanding cellular processes.
  • Existing methods for viscosity measurement can be limited in spatial resolution or applicability to live samples.

Purpose of the Study:

  • To develop a wide-field, time-resolved imaging system capable of simultaneously measuring fluorescence lifetime and rotational correlation time.
  • To demonstrate the system's ability to image variations in solvent viscosity.
  • To apply the technique for probing microviscosity within live cells.

Main Methods:

  • A polarization-resolved imager was employed to capture orthogonal polarization components of fluorescence emission.

Related Experiment Videos

  • A time-gated intensified CCD camera was used for time-resolved detection.
  • The system was validated by imaging fluorescein in a multiwell plate to assess solvent viscosity variations.

    • Main Results:

    • The developed system successfully imaged both fluorescence lifetime and rotational correlation time.
    • Solvent viscosity variations were quantitatively visualized using fluorescein's rotational correlation time.
    • The technique was successfully applied to probe the microviscosity in live cells, demonstrating its biological relevance.

    Conclusions:

    • The novel wide-field time-resolved imaging system offers a powerful tool for quantitative analysis of fluorescence lifetime and rotational dynamics.
    • This technique enables sensitive detection and imaging of microviscosity changes in various environments, including live biological systems.
    • The ability to probe cellular microviscosity opens new avenues for understanding cellular function and disease states.